Team:Ionis Paris/Notebook/18 10 16bis
The overall purpose is to check if our BBX1 biobrick works in E.Coli DH5⍺. This biobrick is composed with XylR coding device with mRFP (Pr-RBS-XylR-RBS-mRFP-Terminator) and has been designed for the monitoring of XylR and to show that this protein is synthesized in the cells when the colonies becomes red thanks to the mRFP protein. BBX1: "XylR coding device with mRFP”, BBa_K2023013 (Pr-RBS-XylR-RBS-mRFP-Terminator). E.Coli DH5⍺ from the -80°C transformed with X1. SOC medium at room temperature. Let the cells on ice 10 minutes for the defrosting. Add SOC medium in order to have a dilution of 1/2, 1/10 and 1/100. Plate the different concentrations (Without dilution and the 3 dilutions) on petri dish with LB Agare + Chloramphenicol at 25 µg/mL final. Expected results: We expected red colonies due to the synthesis of the red protein mRFP. The promoter of this protein is Pr, a strong constitutive promoter. Obtained results: We obtained expected results.
TWe optained red colonies which prove that the mRFP is synthesized by the cells and in the same way the XylR protein.
We have shown that our E.Coli DH5⍺ transformed with the biobrick BBa_K2023013 synthesized XylR in a constitutive way.
XylR Synthesis Test on BBX1
Objectives
Materials
Protocol
Results
Interpretation
